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Mid-Term review group activity (1).docx
Mid-Term Lab Practical Review – Group Activity

1. Match each reagent/tool with its associated technique:

a. Antibiotic filter disc

b. Depression slide

c. Nigrosin

d. Iodine

e. Inoculating loop & agar plate

f. Autoclave

g. Malachite green

h. Immersion oil

i. Bibulous paper

j. Hot water bath

_____ Sterilization

_____ Gram stains

_____ Motility testing

_____ Microscopy with the 100X objective

_____ Kirby-Bauer Test

_____ Isolation streaking

_____ Determination of thermal death time

_____ Negative staining

_____ Endospore staining

_____ Drying your prepared slide

2. Outline the steps for preparing a Gram stain, including preparing the smear.

3. You would expect a Mycobacterium species to be: (Gram +/Acid fast). (pick one)

4. Is Brownian motion the same as true motility? Explain.

Biochemical tests individual (1).docx

BIO2073: Microbiology Lab

Individual Assignment

Biochemical Tests: Phenol Red Fermentation Broth and Catalase Test

1. For the Phenol Red Fermentation Broth, what two key ingredients allow us to test for fermentation? Explain the purpose of each.

2. How can a color change of the broth from red to yellow tell us that fermentation has occurred?

3. What is a Durham tube? Why might we include it in our PR test broth?

4. Write out the equation for the reaction that is occurring during the catalase test.

5. In the catalase test, what is causing the formation of bubbles that we interpret as a positive reaction? (In other words, what is being released during the reaction?)

Bonus! If you tested an organism using the Phenol Red glucose test, and it was positive, could you accurately predict that your organism would also test positive using sucrose? Why or why not?

Antibiotic Sensitivity Lab Report OL.docx

BIO2073- Microbiology Lab

Antibiotic Sensitivity Testing – Group Activity

1. What is the objective of this week’s laboratory exercise?

2. This table was completed using the information from the data file. For each plate, one sample remains to be measured. A ruler is on the slide so that you can measure the zone of inhibition. Use the Interpretive Standards handout to determine the level of resistance: Resistant (R), Intermediate (I), or Sensitive (S).

Antibiotic

Antibiotic Code

Escherichia coli

Proteus mirabilis

Staphylococcus aureus

Zone of inhibition

(mm)

Susceptible/

resistant/ intermediate

Zone of inhibition

(mm)

Susceptible/

resistant/ intermediate

Zone of inhibition

(mm)

Susceptible/

resistant/ intermediate

Bacitracin

B10

6

6

6

Penicillin

P10

6

6

Chloramphenicol

C30

27

20

23

Neomycin

N30

19

20

Erythromycin

E15

10

6

25

Tetracycline

Te30

15

6

22

Streptomycin

S10

15

16

14

Sulfamethoxazole + Trimethoprim

SXT

22

23

3. Based on the results, which antibiotics might be useful against the strain of Proteus mirabilis tested here?

4. The paper discs are labeled with abbreviations (e.g., B10). The letter refers to the name of the antibiotic. What does the number refer to?

Kirby Bauer Individual report.docx

Name: __________________________ Date:____________

BIO2073: Microbiology Lab

Antibiotic Susceptibility Testing: Kirby Bauer Disk Diffusion Method

1. What does the term MIC stand for? Does a lower MIC indicate that a microbe is more or less resistant to an antibiotic?

2. Standardization is critical to obtaining accurate, reliable results with this testing method. Predict what effects each of the following changes might have on the results of the test. (Your lab text has critical information that will help you answer this question.)

a. The agar is only 2 mm deep, instead of the standardized 4 mm:

_________________________________________________________________

___________________________________________________________________________

b. There is visible moisture on the plate: _____________________________________

___________________________________________________________________________

c. The turbidity of your culture was much higher than the 0.5 McFarland standard:

____________________________________________________________________

__________________________________________________________________________

5. Normally, cultures that have been diluted to match the 0.5 McFarland standard must be used within 30 minutes. Why do you think this is important?

______________________________________________________________________________

______________________________________________________________________________

6. How does the antibiotic get from the disk into the agar? (I’m looking for a chemical concept, this can actually be summed up in a single word.) _____________________________

______________________________________________________________________________

7. After incubation, does the antibiotic extend into the agar beyond the zone of inhibition? How does the MIC relate to the zone of inhibition?

______________________________________________________________________________

______________________________________________________________________________

______________________________________________________________________________

______________________________________________________________________________

______________________________________________________________________________

8. Suppose you do this test on a hypothetical Staphylococcus species with the antibiotics penicillin (P 10) and chloramphenicol (C 30). You record zone diameters of 25 mm for both the chloramphenicol and penicillin disks. Which antibiotic would be more effective against this organism? What does this tell you about comparing zone diameters to each other and the importance of the zone diameter interpretive table?

______________________________________________________________________________

______________________________________________________________________________

______________________________________________________________________________

______________________________________________________________________________

______________________________________________________________________________

9. Bonus! Name one other type of test for antimicrobial susceptibility. (1 pt) Explain how this test differs from the Kirby-Bauer test. (1 pt)

Fermentation and Catalase Group (1).docx

Name: __________________________ Date:____________ 

 

BIO2073: Microbiology Lab 

Group Activity

Biochemical Tests: Phenol Red Fermentation Broth and Catalase Test

 

1. Phenol Red Broth: In your Data file, you have results for 8 different unknowns, labeled A-H. Each one has been inoculated into PR broth containing (1) Dextrose (aka, glucose) and (2) sucrose. Record the results in the table below.

Sample

Carbohydrate Source

Result (A/G, A/-, -/-, or K)

Control

Dextrose

Sucrose

A

Dextrose

Sucrose

B

Dextrose

Sucrose

C

Dextrose

Sucrose

D

Dextrose

Sucrose

E

Dextrose

Sucrose

F

Dextrose

Sucrose

G

Dextrose

Sucrose

H

Dextrose

Sucrose

6. If you tested an organism using the Phenol Red glucose test, and it was positive, could you accurately conclude that your organism would also test positive using sucrose instead? Why or why not?

____________________________________________________________________________________

____________________________________________________________________________________

____________________________________________________________________________________

7. a. Write down the equation for the chemical reaction that is occurring during the catalase test:

b. In the catalase test, the formation of bubbles is considered a positive result, indicating the organism produces the enzyme catalase. What product of the reaction is responsible for the formation of bubbles?

____________________________________________________________________________________

8. Name a medically significant microbe that would test positive for coagulase:

_________________________________________________________________

*Bonus question on next page!

Bonus! Imagine that your coagulase test was negative with the slide test, but positive for the tube test. What is the most likely explanation for this observation? (Assume that we can rule out human error.)

____________________________________________________________________________________

____________________________________________________________________________________

____________________________________________________________________________________

Differential II VR group (1).docx

BIO2073: Microbiology Lab

Differential Tests II – Group Activity

Part A: Litmus Milk

Review the results in the PowerPoint file “Litmus Milk Data” and answer the following questions:

1. Based on the Day 2 results, give your best interpretation for each sample:

Sample

Litmus Color

Ferments Lactose?

A

B

C

D

E

F

G

H

2. Based on the Day 7 Results, give your best interpretation for each sample.

Sample

Clot formation?

Gas Production?

Reduction of Litmus?

A

B

C

D

E

F

G

H

Part B. EnteroPluri Data

For each sample, give your best interpretation of the results and generate a 5-digit Biocode. Then use the “EnteroPluri” PDF file to look up your number and determine the identify of your organism. Indole results have been provided for you.

Sample 1:

Sample 2:

Sample 3:

Sample 4:

Eukaryotic microscopy individual activity (4).docx

BIO2073: Microbiology Lab

Eukaryotic Microscopy: Individual Assignment

Several of the organisms we looked at cause disease in humans. Pick
3
of the diseases listed below, and for each of them, provide:

a. The name of the causative organism

b. The body system(s) that it affects

c. Its symptoms

d. How it’s prevented and/or treated

· Aspergillosis

· Malaria

· Trichomoniasis

· African sleeping sickness

· Amoebiasis

· Candidiasis

· Taeniasis

Please be sure to include this information in your own words and cite your sources appropriately in APA format.

Heat Lab OL.docx

BIO2073- Microbiology lab

Summer Quarter 2020

Lab Report: Heat as a method of control

1. What was the objective of this lab exercise?

 

2. Use the following table to document your data. For each plate that you look at, be sure to indicate the species, and the temperature used. Then, for each time point, evaluate the relative amount of growth in each section using a scale from 0-4 where
0 = No growth 4= heaviest growth

Organism

63°C

72°C

0 sec

30 sec

1 min

2 mins

5 mins

15mins

0 sec

30 sec

1 min

2 mins

15 mins

3. Why was it important to plate out a sample at 0 seconds, prior to exposing the culture to heat?

4. In general, was 63°C or 72°C more effective at killing the microbes? Does this align with the results you expected? Why or why not?

4. Both “young” (less than 24 hours) and “old” (several weeks) cultures of B. subtilis were used in this experiment. Why do you think this was done? Hint: what do you know about the Bacillus genus and their survival mechanisms? Why use Bacillus and not old/young E.coli for example?

6. The thermal death time and the term death point are two different ways to describe a microbe’s sensitivity to heat. Which of these two measurements were we evaluating in this experiment? Explain your answer.

7. Give an example of a non-laboratory use of each of the following methods to control microbial growth:

a. Incineration:

b. Pasteurization:

c. Autoclaving:

Bonus! The species Geobacillus stearothermophilus is often used as an indicator species to test the effectiveness of autoclaving. Why do you think this species is used instead of E. coli or B. subtilis?

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